Biopolymer Drug Loading Method

ABSTRACT

A method of producing a drug-loaded Poly (Glycerol Sebacate) (PGS) comprising providing PGS; dissolving at least one drug in a solvent; incubating the PGS in the solvent; and evaporating the solvent.

CROSS-REFERENCE TO RELATED APPLICATION(S)

This application claims the benefit of U.S. Provisional Application No.62/130,180, filed on Mar. 9, 2015; which is incorporated herein byreference in its entirety

FIELD OF THE INVENTION

Disclosed herein are methods for improved drug loading of thebio-polymer Poly (Glycerol Sebacate) (PGS).

BACKGROUND OF THE INVENTION

Drug loaded bio-polymer implants provide advantages in many tissueengineering applications. For example, bio-polymer implants loaded withantibiotics can reduce risk of bacterial biofilm formation that can leadto implant site infection, immune reactions, implant failure, andbacteria-associated systematic problems. Poly (Glycerol Sebacate) (PGS)is a recently developed is a biocompatible, biodegradable, elasticpolymer that has been widely tested for many tissue engineeringapplications. However, current PGS drug loading methods have a number oflimitations. For example, past methods perform drug loading at the PGSpre-polymer stage. During the cross-linking process, residual solventand loaded drug may affect the condensation proliferation process. Thisin turn may alter crosslinking speed and condition, crosslink drugand/or solvent into the polymer chain and alter drug formation, leadingto reduced drug activity or to increased toxicity. Furthermore, the highheat required during crosslinking of pre-PGS can lead to degradation ofmany antimicrobial drugs. Accordingly, there is a need in the art for animproved method of PGS drug loading.

BRIEF SUMMARY OF THE INVENTION

Disclosed herein is a method of producing a drug-loaded Poly (GlycerolSebacate) (PGS) comprising providing PGS, dissolving at least one drugin a solvent, incubating the PGS in the solvent; and evaporating thesolvent. In certain aspects, the step of providing PGS further comprisessynthesizing PGS by the steps of providing a mixture of gylceraol andsebacic acid, heating the mixture, polymerizing the mixture in a vacuumoven in N2 atmosphere to form a pre-polymer, cross-linking the PGSpre-polymer to form PGS, and balancing the PGS. According to certainaspects, the PGS is balanced in phosphate buffered saline (PBS). In yetfurther embodiments, PGS is cut into a desired form prior to incubationin the drug containing solvent.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a flow chart of the disclosed drug loading method, accordingto certain embodiments.

FIG. 2 shows PGS drug loading efficiency under various ethanolconcentrations (A) and temperatures (B), according to certainembodiments.

FIG. 3 shows daily (A) and accumulated (B) release profile of Berberinein B-PGS and BC-PGS, according to certain embodiments.

FIG. 4 shows Young's modulus (A) and strain at break (B) of untreatedPGS (Untreated), drug loading treated PGS (PGS), berberine loaded PGS(B-PGS), chlorhexidine loaded PGS (C-PGS) and berberine-chlorhexidineloaded PGS (BC-PGS), according to certain embodiments.

FIG. 5 shows surface wettability data for PGS, B-PGS, C-PGS and B—C-PGS,according to certain embodiments.

FIG. 6 shows antimicrobial data for S. aureus (A), MRSA (B) and E. coli(C), according to certain embodiments.

FIG. 7 shows SEM images of sample surfaces before (A˜D) and afterculture with S. aureus (E˜H) and E. coli (I˜L) for 3 days, according tocertain embodiments.

FIG. 8 shows fluorescence images of HGF-1 cells on the surface of (A)PGS, (B) B-PGS, (C) C-PGS and (D) BC-PGS after three days of seeding,according to certain embodiments.

DETAILED DESCRIPTION

Although the present invention has been described with reference topreferred embodiments, persons skilled in the art will recognize thatchanges may be made in form and detail without departing from the spiritand scope of the invention.

Ranges can be expressed herein as from “about” one particular value,and/or to “about” another particular value. When such a range isexpressed, a further aspect includes from the one particular valueand/or to the other particular value. Similarly, when values areexpressed as approximations, by use of the antecedent “about,” it willbe understood that the particular value forms a further aspect. It willbe further understood that the endpoints of each of the ranges aresignificant both in relation to the other endpoint, and independently ofthe other endpoint. It is also understood that there are a number ofvalues disclosed herein, and that each value is also herein disclosed as“about” that particular value in addition to the value itself. Forexample, if the value “10” is disclosed, then “about 10” is alsodisclosed. It is also understood that each unit between two particularunits are also disclosed. For example, if 10 and 15 are disclosed, then11, 12, 13, and 14 are also disclosed.

References in the specification and concluding claims to parts by weightof a particular element or component in a composition denotes the weightrelationship between the element or component and any other elements orcomponents in the composition or article for which a part by weight isexpressed. Thus, in a compound containing 2 parts by weight of componentX and 5 parts by weight component Y, X and Y are present at a weightratio of 2:5, and are present in such ratio regardless of whetheradditional components are contained in the compound.

A weight percent (wt. %) of a component, unless specifically stated tothe contrary, is based on the total weight of the formulation orcomposition in which the component is included.

As used herein, the terms “optional” or “optionally” means that thesubsequently described event or circumstance can or cannot occur, andthat the description includes instances where said event or circumstanceoccurs and instances where it does not.

Disclosed herein is a method of producing a drug-loaded Poly (GlycerolSebacate) (PGS) comprising providing PGS, dissolving at least one drugin a solvent, incubating the PGS in the solvent; and evaporating thesolvent. According to certain embodiments, providing PGS furthercomprises synthesizing PGS by the steps of providing a mixture ofgylceraol and sebacic acid, heating the mixture, polymerizing themixture in a vacuum oven in N₂ atmosphere to form a pre-polymer,cross-linking the PGS pre-polymer to form PGS, and balancing the PGS.According to certain embodiments, the PGS is balanced in phosphatebuffered saline (PBS). In yet further embodiments, PGS is cut into adesired form prior to incubation in the drug containing solvent.

In certain exemplary embodiments, gylceraol and sebacic acid are addedin approximately equimolar amounts.

In certain embodiments, the mixture is polymerized for about 24 hours.In further embodiments, the mixture is polymerized for between about 8hours to about 36 hours.

In still further embodiments, the pre-PGS is crosslinked for about 72hours. According to yet further embodiments, the pre-PGS is crosslinkedfrom about 12 hours to about 144 hours.

According to certain embodiments, the solvent is a low toxicity solvent.In further embodiments, the solvent is ethanol. In yet furtherembodiments, ethanol is about 100% ethanol. In still furtherembodiments, the solvent is water. In further embodiments, the solventis acetone, DMSO, or 2,2,2-trifluoroethanol. One skilled in the art willappreciate that other organic solvents may be used.

In certain embodiments, incubating the PGS in the solvent is performedbetween the temperatures of about −20° C. to about 100° C. In furtherembodiments, incubating the PGS in the solvent is performed between thetemperatures of about −114° C. (melting point of ethanol) to 78° C.(boiling point of ethanol). In yet further embodiments, incubating thePGS in the solvent is performed at about 37° C.

According to certain embodiments, the at least one drug is anantimicrobial. In further embodiments, the antimicrobial ischlorhexidine. In yet further embodiments, the antimicrobial isberberine. In still further embodiments, the antimicrobial is berberineand chlorhexidine. In still further embodiments, the antimicrobial isselected from a group consisting of: penicillin, streptomycin,rifampicin, levofloxacin, tetracycline, erythromycin, and triclosan,According to further embodiments, the disclosed methods can be used toload other organic solvent and water soluble drug(s), includinganti-cancer and anti-inflammatory drugs.

Disclosed herein is a drug-loaded PGS product prepared by a processcomprising the steps of providing PGS; dissolving at least one drug in asolvent; incubating the PGS in the solvent; and evaporating the solvent.According to certain embodiments, providing PGS further comprises thesteps of providing a mixture of gylceraol and sebacic acid; heating themixture; polymerizing the mixture in a vacuum oven in N₂ atmosphere toform a pre-polymer; cross-linking the PGS pre-polymer to form PGS; andbalancing the PGS.

According to certain embodiments, the at least one drug of the productproduced by the disclosed process is an antimicrobial. In furtherembodiments, the antimicrobial is chlorhexidine. In yet furtherembodiments, the antimicrobial is berberine. In still furtherembodiments, the antimicrobial is berberine and chlorhexidine. In stillfurther embodiments, the antimicrobial is selected from a groupconsisting of: penicillin, streptomycin, rifampicin, levofloxacin,tetracycline, erythromycin, and triclosan, According to furtherembodiments, the disclosed methods can be used to load other organicsolvent and water soluble drug(s), including anti-cancer andanti-inflammatory drugs.

According to certain embodiments, the product disclosed herein can beused for a variety of tissue engineering purposes. For example,according to certain embodiments, the disclosed drug-loaded PGS productcan be used for cardiac tissue engineering, vascular tissue engineering,cartilage tissue engineering, retinal tissue engineering and nervetissue engineering. Depending on loaded drug(s), the drug loaded PGS canexhibit multiple biological activities while retaining its originalmechanical and biological properties.

In certain embodiments, drug loading of PGS according to the disclosedprocess does not affect the mechanical properties of PGS. For example,PGS elasticity of drug loaded PGS, according to certain embodiments, issubstantially similar to non-drug loaded PGS.

According to still further embodiments, the disclosed drug-loaded PGSproduct is capable of sustained drug release. In still furtherembodiments, the disclosed drug-loaded PGS performs sustained drugrelease for at least 60 days.

According to certain exemplary embodiments, the disclosed drug-loadedPGS product has antimicrobial properties. In still further embodiments,the disclosed drug-loaded PGS product reduces bacterial adhesion. In yetfurther embodiments, the disclosed drug-loaded PGS product has highcompatibility with human cell types. In still further embodiments, thedisclosed drug-loaded PGS product provides an effective treatment forperiodontal disease when administered as a dental implant.

Experimental

The following examples are put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of certainexamples of how the compounds, compositions, articles, devices and/ormethods claimed herein are made and evaluated, and are intended to bepurely exemplary of the invention and are not intended to limit thescope of what the inventors regard as their invention. However, those ofskill in the art should, in light of the present disclosure, appreciatethat many changes can be made in the specific embodiments which aredisclosed and still obtain a like or similar result without departingfrom the spirit and scope of the invention.

Materials and Methods

S. aureus (ATCC 6538), E. coli (ATCC 15597) and Methicillin-resistantStaphylococcus aureus (MRSA, ATCC BAA-811) were purchased from AmericanType Culture Collection (ATCC, Manassas, Va.). Human gingival fibroblast(HGF-1) were purchased from American Type Culture Collection (ATCC,Manassas, Va.). All reagents used in our work including glycerol,sebacic acid, berberine chloride, chlorhexidine and ethanol were fromSigma-Aldrich (Sigma-Aldrich, St Louis, Mo.) unless otherwise specified.

Synthesis of PGS

Poly (Glycerol Sebacate) (PGS) was synthesized following the disclosedmethod, shown schematically in FIG. 1. Briefly, equimolar of glyceroland sebacic acid were melted down and mixed evenly. The mixture was thenput into vacuum oven (Fisher Isotemp Vacuum Oven Model 281A, FisherScientific, Waltham, Mass.) at 120° C. in N2 atmosphere for 24 hours forthe synthesis of PGS pre-polymer (pre-PGS). Pre-PGS was thencross-linked at 120° C. under vacuum for 72 hours to make the 3-Dcross-linked PGS elastic polymer. Synthesized PGS was then cut into 10mm×10 mm×4 mm cubes and balanced in phosphate buffered saline (PBS) for3 days before use.

Swelling Drug Loading

Due to its unique 3-D structure, PGS does not get dissolved in mostsolvents. However, it swells in many organic solvents, and that propertycan be used for drug loading. As will be appreciated by those skilled inthe art, many drugs can be loaded into PGS using various solvents(organic solvents or water). In the instant examples, berberine andchlorhexidine were selected as exemplary drugs and ethanol as anexemplary drug-loading solvent. One skilled in the art will appreciatethat other drugs and solvents can be used.

Selected drugs (individually and together) were dissolved 100% ethanolto make a saturated solution. PGS cubes were then put into saturateddrug/ethanol solution for swelling and drug loading for 2 days. Drugloaded PGS cubes were slowly dried and cleaned afterward. In the presentexample, berberine loaded PGS (B-PGS) cubes, chlorhexidine loaded PGS(C-PGS) cubes, and berberine-chlorhexidine loaded PGS (B-C-PGS) cubeswere prepared with saturated drug ethanol solution respectively and PGScubes treated with pure 200 ethanol were employed as control.

PGS Swelling Test

PGS was synthesized, buffered and cut into 10 mm×10 mm×4 mm cubes asmentioned above. PGS cubes were weighted (W_0) before swelling test.Ethanol and acetone were selected for swelling test because they arecommonly used, less toxic class 3 solvents in pharmaceutical industry.PGS cubes were weighted again (W_1) after 48 hours drug loadingtreatment. The swelling behavior (SB_(w)(%)) was calculated based onweight change with the following equation:

${{SB}_{w}(\%)} = {\frac{W_{1} - W_{0}}{W_{0}} \times 100\%}$

PGS swelling behavior in di water and phosphate buffered saline (PBS)were also tested for reference.

Drug Loading and Release Test

Drug loaded PGS cubes were placed into 2,2,2-trifluoroethanol (TFE) forthree days for a complete release. Berberine concentration wasdetermined by measuring the optical density of the supernatant at 344 nmusing a micro-plate reader (Tecan infinite M200, Tecan Group Ltd.Männedorf, Switzerland). Two drug loading parameters—temperature andethanol/water ratio were optimized for higher drug loading efficiency.All tests were performed at 37° C. with 100% ethanol unless otherwisespecified.

Sterile PBS was used for drug release test. One cube of treated PGSsample was placed into a glass vial containing 10 ml sterile PBS. Theglass vial was then placed into a mechanical to test the drug releasingproperty at 37° C., 150 rpm. Sterile PBS was changed daily and releaseddrug was measured using the same method as in drug content test.

Mechanical Test

Synthesized PGS were cut into 50 mm×10 mm×4 mm strips. Drug(s) wereloaded into PGS strips using the same method while an untreated groupwas prepared without any drug loading treatments. For the mechanicaltest, five groups of samples (PGS, B-PGS, C-PGS, BC-PGS and untreated)were mounted on the sample loader of MTS insight electromechanicaltesting system (MTS systems corporation, Eden Praire, Minn.). Young'smodulus and strain at break were measured by running MTS tensile methodwith five groups of samples.

Surface Wettability Test

Surface wettability was determined by contact angle goniometry by usingVCA optima contact angle analysis system (AST products, Inc., Billerica,Mass.). In this test, one drop (2 μL) of distilled water was droppedonto the horizontal surface of a sample cube. Photos were taken usingthe contact angle analysis system after 1 minute and contact angles weremeasured with its software workstation.

Antimicrobial Test

General antimicrobial test was taken following ASTM standard E2149-10.E. coli was cultured in LB broth (Lennox, Fisher Scientific, Pittsburgh,Pa.) overnight. S. aureus and MRSA was cultured in tryptic soy broth(Fluka analytical, Sigma-Aldrich, St Louis, Mo.) overnight. All bacteriawere grown to OD470=0.30˜0.35 and was further diluted 100 times withPBS. PGS, B-PGS, C-PGS and BC-PGS cubes were put into 20 ml dilutedbacteria solution and shaken vigorously in an isotemp shaker (MAXQ 4450,Thermo Scientific, Waltham, Mass.) at 37° C. The colony formation unit(CFU) of the supernatant was tested after 1 and 4 hours to determine theantimicrobial activity of four groups of samples.

Bacteria Adhesion Test

Gram-positive bacteria S. aureus (ATCC 6538) and gram negative bacteriaE. coli (ATCC 15597) were used for bacteria adhesion test. S. aureus wasgrown in tryptic soy broth, and E. coli was grown in Luria-Bertani (LB)broth overnight ahead of use. Both bacteria were then diluted to 105CFU/ml using the corresponding broth. Four groups of PGS cubes (PGS,B-PGS, C-PGS and B—C-PGS) were immersed in 20 ml of diluted bacteriabroth individually and cultured at 37° C. for 3 days.

Samples were taken out and washed thoroughly with PBS after 3 days ofculture. To test the bacteria adhesion, samples were fixed in 2.5%glutaraldehyde for 2 h and dehydrated with a series of ethanol solution(70%, 80%, 90%, and 100%) for 10 minutes each. Samples were then driedand coated with 10 nm gold with a sputter coater (Cressington 108,Cressington Scientific Instruments Ltd. Watford, UK) and observed with aScanning Electron Microscope (SEM) (FEI Quanta 450, FEI Company,Hillsboro, Oreg.).

Cell Compatibility Test

Human gingival fibroblast cells (HGF-1, ATCC® CRL-2014™) were used forthe cell compatibility assay. HGF-1 cells were cultured in DMEM/HighGlucose supplemented with 1% non-essential amino acids (FisherScientific, Pittsburgh, Pa.), 1% antibiotics (Fisher Scientific,Pittsburgh, Pa.), and 10% fetal bovine serum (Sigma-Aldrich, St Louis,Mo.). Four groups of samples were sterilized by UV and were balanced incell culture media for 7 days in a 24 well plate before the test. HGF-1cells were then seeded onto the surface of the membranes at aconcentration of 100,000 cells per well. PGS cubes were taken out andfixed with 2.5% glutaraldehyde at 4° C. overnight. Samples were thenpermeabilized with 0.5% triton X-100 for 10 minutes at room temperature.Permeabilized cells were stained with 100 nM rhodamine phalloidin(Molecular Probes Life Technologies, Eugene, Oreg.) in the dark at roomtemperature for 30 minutes. After rhodamine phalloidin stain, sampleswere treated with 0.3% (w/w) Sudan Black for 30 minutes to reducebackground emission. Fluorescence images were taken by Axiovert 200fluorescence microscope (Axiovert 200, Carl Zeiss AG, Jena, Germany).

Statistical Analysis

All quantitative tests were carried out with three parallel samples,with results presented in the form of mean±standard deviation (S.D.)unless otherwise specified. The statistical significance of the dataobtained was analyzed by one-way ANOVA. Probability values of p<0.05were interpreted as statistically significant.

Results PGS Swelling and Drug Loading

The synthesized PGS is a transparent, elastic polymer. PGS is able toswell in many organic solvents and water (Table 1). Ethanol was chosenas drug loading solvent because: 1) it has low toxicity; 2) the twomodel drugs used—berberine and chlorhexidine—both have good solubilityin ethanol; and 3) PGS can maintain its surface morphology after ethanoltreatment. As shown in FIG. 2 (panel A), the solvent selection testshowed that berberine loading efficiency decreases with the decrease ofethanol composition in the solvent system and 100% ethanol shows highestberberine loading efficiency.

TABLE 1 Solvent Water PBS Ethanol Acetone Swelling 3.82% ± 3.41% ±55.22% ± 113.98% ± 0.25% 0.19% 0.51% 1.95%

As shown in FIG. 2 (panel B), increasing temperature is also able toincrease berberine loading efficiency. However, drugs can be unstable inhigh temperature and 37° C. was chosen as drug loading temperature as abalance of sufficient drug loading efficiency and drug stability.

Using tested PGS swelling behavior (SB_(w)), drug loading concentration(c₁) and solvent density (ρ), expected drug loading (EDL (w/w)) can becalculated with the following concentration:

${{EDL}\left( {w/w} \right)} = {\frac{{SB}_{w}}{\rho} \times c_{l} \times 100\%}$

The expected drug loading is calculated by assuming that drugs andmaterial have no interaction while the only mechanism for drug loadingis the swelling process. However, as shown in Table 2, tested drugloading is higher than expected drug loading for both berberine andchlorhexidine. This indicates that PGS produces a positive effect on theberberine and chlorhexidine loading process.

TABLE 2 Expected and actual drug loading efficiency of B-PGS and C-PGSExpected Tested Drug Loading Drug Loading Drug Loading EfficiencyBerberine 0.402% 0.699% 173.71% in B-PGS Chlorhexidine 0.019% 0.042%221.49% in C-PGS

Releasing Behavior of Drug Loaded PGS

Drug release was assessed in sterile PBS to avoid PGS degradation. Thedaily and accumulated release curve of berberine in B-PGS and BC-PGS isshown on FIG. 3. 15% of berberine was released after 62 days with astable daily release concentration of around 1 μg/ml. Chlorhexidinerelease concentration was too low to be detected with the detectionmethod (<5 μg/ml). The results indicate that drug-loaded PGS performedsustained release of the drugs.

Mechanical Test

One important property of PGS is its elastic property. To determinewhether drug loading affected mechanical properties of PGS, anelongation test was performed. As shown in FIG. 4, PGS (drug loadingtreated), B-PGS, C-PGS and BC-PGS showed no significant differencecompared with untreated (untreated PGS) samples as measured by Young'smodulus (FIG. 4, panel A) and strain at break (FIG. 4, panel B). Thisresult indicated the disclosed swelling drug loading method and loadeddrug(s) did not change the mechanical property of original PGS.

Surface Wettability Test

Material surface wettability is an important property for tissueengineering. Surface wettability of PGS, B-PGS, C-PGS and BC-PGS wastested using water contact angle test, as shown in FIG. 5. The contactangle of PGS is 53.5°, which is suitable for cell attachment.Hydrophilic drug berberine decreased water contact angle to 46°. Thisindicated an improved surface wettability and can have some beneficialin tissue engineering. On the other side, hydrophobic drug chlorhexidineincrease water contact angle to 83.5°, making the surface of C-PGS morehydrophobic. Interestingly, when hydrophilic drug berberine andhydrophobic drug chlorhexidine loaded together, BC-PGS performed asimilar water contact angle (47.7°) as B-PGS.

Antimicrobial Test

FIG. 6 shows the result of general antimicrobial test against of S.aureus (panel A), MRSA (panel B) and E. coli (panel C) for PGS, B-PGS,C-PGS and BC-PGS. (Star sign indicates the marked group has significantdifference to PGS group of each time point. * p<0.05, ** p<0.01.)Chlorhexidine loaded groups (C-PGS and BC-PGS) showed good antimicrobialeffect against typical gram-positive bacteria S. aureus, typicalgram-negative bacteria E. coli and typical antibiotic-resistant bacteriaMRSA.

C-PGS killed 99.86% S. aureus, 94.36% E. coli, and 99.96% MRSA whileBC-PGS killed 99.88% S. aureus, 96.13% E. coli and 100% MRSA after 4hours of incubation. B-PGS does not show any significant antimicrobialeffect in general antimicrobial test. However, in most test groups,BC-PGS showed higher antimicrobial effect than C-PGS.

Bacteria Adhesion of Drug Loaded PGS

As shown in FIG. 7, SEM images showed that PGS samples had a smooth,clear surface with occasionally micro-scaled crests and troughs (FIG. 7panel C) as well as deposited drug crystal (FIG. 7 panel D).

After incubating with S. aureus for 3 days, significant amounts ofbacteria was observed on the surface of PGS (FIG. 7, panel E) whilefewer bacteria was found on C-PGS (FIG. 7, panel G). Interestingly,although B-PGS had weaker antimicrobial effect than C-PGS, fewerbacteria can be found on its surface (FIG. 7, panel F). In addition,very few bacteria were observed on the surface of B—C-PGS (FIG. 7, panelH), indicating an excellent anti-adhesion property against S. aureus.

Different result was observed on the E. coli co-cultured groups. Theattached bacteria concentration was less than S. aureus on all samplesurfaces after 3-day incubation. C-PGS group showed better anti-adhesioneffect than B-PGS group against E. coli. Moreover, very few bacteriawere found on the surface of B-C-PGS.

Cell Compatibility Test

Human gingival fibroblast (HGF-1) was used because it is the mostabundant cell in periodontal soft tissue. FIG. 6 shows the fluorescenceimages of HGF-1 cell grown on sample surfaces after 3 days. Compared tothe PGS group (FIG. 8, panel A), more cells were found on the surface ofB-PGS (FIG. 8, panel B) and BC-PGS (FIG. 8, panel D). While fewer cellswere found on the surface of C-PGS (FIG. 8, panel C). This showsberberine loaded PGS groups have improved cell compatibility with HGF-1.

Although the present invention has been described with reference topreferred embodiments, persons skilled in the art will recognize thatchanges may be made in form and detail without departing from the spiritand scope of the invention.

What is claimed is:
 1. A method of producing a drug-loaded Poly(Glycerol Sebacate) (PGS) comprising: a. providing PGS; b. dissolving atleast one drug in a solvent; c. incubating the PGS in the solvent; andd. evaporating the solvent.
 2. The method of claim 1, wherein providingPGS further comprises: a. providing a mixture of gylceraol and sebacicacid; b. heating the mixture; c. polymerizing the mixture in a vacuumoven in N₂ atmosphere to form a pre-polymer; d. cross-linking the PGSpre-polymer to form PGS; and e. balancing the PGS.
 3. The method ofclaim 2, wherein the mixture is polymerized for about 24 hours.
 4. Themethod of claim 2, wherein the pre-PGS is cross-linked for about 72hours.
 5. The method of claim 1, wherein the solvent is a low toxicitysolvent.
 6. The method of claim 5, wherein the solvent is ethanol. 7.The method of claim 1, wherein the incubating the PGS in the solvent isperformed at between the temperatures of from about −20° C. to about100° C.
 8. The method of claim 7, wherein the incubating the PGS in thesolvent is performed at about 37° C.
 9. The method of claim 1, whereinthe solvent is evaporated at room temperature for at least 7 days. 10.The method of claim 1, wherein the at least one drug is anantimicrobial.
 11. The method of claim 10, wherein the antimicrobial ischlorhexidine.
 12. The method of claim 10, wherein the antimicrobial isberberine.
 13. The method of claim 10, wherein the antimicrobial isberberine and chlorhexidine.
 14. The method of claim 10, wherein theantimicrobial is selected from a group consisting of: penicillin,streptomycin, rifampicin, levofloxacin, tetracycline, erythromycin, andtriclosan.
 15. A drug-loaded Poly (Glycerol Sebacate) (PGS) productprepared by a process comprising the steps of: a. providing PGS; b.dissolving at least one drug in a solvent; c. incubating the PGS in thesolvent; and d. evaporating the solvent.
 16. The drug-loaded PGS ofclaim 15, wherein providing PGS further comprises: a. providing amixture of gylceraol and sebacic acid; b. heating the mixture; c.polymerizing the mixture in a vacuum oven in N₂ atmosphere to form apre-polymer; d. cross-linking the PGS pre-polymer to form PGS; and e.balancing the PGS.
 17. The drug-loaded PGS of claim 15, wherein the atleast one drug is an antimicrobial.
 18. The drug-loaded PGS of claim 17,wherein the antimicrobial is chlorhexidine.
 19. The drug-loaded PGS ofclaim 17, wherein the antimicrobial is berberine.
 20. The drug-loadedPGS of claim 17, wherein the antimicrobial is selected from a groupconsisting of: penicillin, streptomycin, rifampicin, levofloxacin,tetracycline, erythromycin, and triclosan.